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3,545,925 Patented July 17, 1962 Fine 3,045,025 BIS QUATERNARY OXIMES Brennie E. Hackley, Jr., and Edward J. Poziomek, Edgewood, and George M. Steinberg, Baltimore, MIL, assignors to the United States of America as represented by the Secretary of the Army No Drawing. Qriginal application Apr. 28, 1959, Ser. No. 809,578. Divided and this application Get. 11, 1961, Ser. No. 151,121

' 1 Claim. (Cl. 260-296) (Granted under Title 35, U.S. Code (1952), sec. 266) This invention described herein may be manufactured and used by or for the Government of the United States of America for governmental purposes without the payment to us of any royalty thereon.

This is a division of application Serial No. 809,578, filed April 28, 1959.

This invention is directed to certain diquaternary pyridinium halide oximes which are useful as chemotherapeutic and prophylactic agents for mammals poisoned by anticholinesterases, particularly the nerve gas known as GB or sarin, i.e., isopropyl methylphosphonofiuoridate.

The invention relates to l,l-polymethylene bis (4-forniylpyridinium) halide dioximes wherein the polymethylene group contains from 2 to 6 carbon atoms. These compounds have the structural formula N+--R N+2X- O CH=NOH CH=NOH (I) where R is a polymethylene group having from two to six carbon atoms and X is chloride, bromide or iodide.

This invention further includes compounds of Formula I, but in which R is a polymethylene group containing from seven to ten carbon atoms.

This invention further includes 1,l'-(2,5 dimethyl-pphenylenedimethylene) bis (4-formylpyridinium) halide dioximes of the formula CH=NOH OH=NOH 1 1 The invention also relates to 1,1'-(2-butenylene)' bis (4-formylpyridinium) halide dioximes of the formula C H=NO H CH=NO E (III) The invention further relates to '1, 1 -polymethylene bis (3-formylpyridinium) halide dioximes of the formula NiloH=Non CH=NOH v I (I Va) wherein R is a polymethylene group containing fi-om 2 to 6 carbon atoms.

Another class'of compounds included are the 1,1'-(p- 7 many insecticides.

2 phenylenedimethylene) bis (3-formylpyridinium) halide dioxirnes of the formula OBI=NOH OH=NOH (IVb) This invention further relates to unsymmetrical bisquaternary 4-formylpyridinium halide monoximes of the formula CH=NOH wherein R" represents either. three lower alkyl groups the hydrocarbon portion of the pyridine ring.

This invention also relates to the l,1-trimethylene bis (3-amidocximopyridinium) halides N+(OHg) -N.+2X-

c 1=Non (|3=NOH NH; N Hz This invention also relates to the compounds (VII) The invention also relates to the compounds N+ (CH3) 3N+2X C H=N O H i (VIII) While all these compounds are useful in varying degree for the purposes set out above, the diiierent groups exhibit quite striking differences in effectiveness. In all the above formulas, X is chloride, bromide or iodide, which appear to be equivalents as to physiological action, except for their effect on solubility.

Wilson et al., in Patent No. 2,816,113, granted Dec. 10, 1957, disclose a group ofcompounds Which'are effective as antidotes for treatment of mammals poisoned by compounds of high anticholinesterase activity, suchas the nerve gases diisopropylphosphorofluoridate (DFP), isopropyl methylphosphonofluoridate (GB), and O-ethyl, N,N-clirnethyl phosphoroamidocyanidate (GA), as well as other related organic phosphorus compounds, including The compound of the Wilson et a1. group which has received by far the most attention. is

,2-formyl-l-methyl 'pyridiniurn iodide oxime, commonly which had been poisoned with paraoxon and then treated with Z-PAM.

Nerve gas poisoning has been treated symptomatically with drugs which are pharmacologically' antagonistic to acetyl choline. Such a compound is atropine and it is at present the recommended remedy. Recently, 2-PAM has been reported to enhance considerably the activity of atropine in the chemotherapeutics of poisoning due to organophosphorus compounds.

The compounds of Formula I above in which R contains from 2 to 6 carbon atoms are appreciably more effective than 2-PAM as reactivators of GB-inhibited acetylchlorinesterase and also in enhancing the activity of atropine in both therapy and propylaxis. When X- is bromide, the variation of the rate constant for the in vitro reactivation of GB inhibited eel acetylcholinesterase at pH 7.4 and 25 C. was found to be as follows:

When administered in combination with atropine to animals poisoned with GB the order of effectiveness was somewhat different. Under these conditions the compound in which R: (CH i.e., l,1-tn'methylene bis (4- formylpyridinium ibromide dioxime also known as TMB-4, was most efiective. In rats challenged with a 2 LD dose of GB administered intervenously, all of a group of six animals survived if the atropine-TMB-4 combination was administered intravenously immediately after poisoning. The atropine-Z-PAM combination saved only two of the group of animals. On the other hand, with dogs which were given a 20 LD dose of GB subcutaneously the survival ratios were the same (4/5) for the two treatments, which were given intravenously when symptoms appeared. However, the recovery time was much shorter for the surviving animals which received the TMB-4, i.e., 2 hours, as against 24 hours for those receiving the 2-PAM.

A summary of the reactivation rates and survival ratios for these compounds when administered therapeutically to rats together with atropine is as follows:

group.

Our compounds may be employed prophylactically, i.e., injected before exposure to the anticholinesterase agent, e.g., GB, or therapeutically, i.e., injected subsequent (to exposure.

The following series of experiments compares the effectiveness of our presently preferred compound TMB-4, with Z-PAM applied to various animals by these two methods. The animals were poisoned by injections of GB as follows.

Mice 0.173 mg./l g. (LD Rats 0.126 mg./kg. (2 LD Rabbits:

Intravenous 0.340 mg./kg. (20 LD Subcutaneous 0.900 mgjkg. (20 LD Dogs and cats:

Intravenous 0.440 mg./kg. (20 LD Subcutaneous 0.900 mg./kg. (20 LD Mg./kg.

l Rats 4 Rabbits 2 Dogs and cats 0.5

The prophylactic doses were given within two minutes prior to the injection of the GB, the therapeutic doses 18 so soon as poisoning symptoms were visible.

Table 3 shows the results.

Table 3 A. PROPHYLACTIC 20 2-PAM TMB4 Animals Survival Ratio Survival Ratio Drils zc Dolsf emg. g. 20 Without With Without With Atropine Atropine Atropine Atropine The recovery periods, i.e., time for disappearance of 40 symptoms of poisoning, among survivors in the above tests,

with atropine, were as follows.

Table 4 2-PAM TMB-4 Animals Prophylactic Therapeutic Prophylactic Therapeutic Rats 60 min 15 min.

Rabbits 3 hr 30 min.-- 2 hr. OM 5 hr 5 hr- 5 hr. 24 hr Dogs 24 hr 24 hr 1% hr 3 hr.

The compounds of Formula I in which R contains from 7 to 10 carbon atoms are less efliective than those of our preferred group. For these compounds the reactivation rate constant and the survival ratio for rats (measured as given above) were as follows, X- being bromide.

Table 5 R Rate Survival Constant Ratio (GB) 2X10 0/4 1. 2X10 0/4 0/4 1. 4X10 0/6 termediate those of the two subgroups of Formula I. When X" was chloride the compound had the following properties. (In this and all following tables the survival ratios are those for rats challenged by 2 LD doses of 6 PREPARATION OF COMPOUNDS 4-pyridinecarboxaldehyde oxime was prepared by warming on a steam bath a neutralized aqueous solution of 4-pyridinecarboxa1dehyde and hydroxylamine hydrofiy i (VX) and the Oxlme was employed therapeutl' 5 chloride. The oxime had a melting point of 1301 30 .5

Compounds of Formula III showed reactivation rates 5 53: 2 and 3 oxlmes were produced by snmlar aasaiata :5assiststaaafiaaa h to w stant was 8x10 as compared to the value for the 10 out by reactmg th? proper omme wlfl-l a lm-dlhaloalkane R: ZM member of our preferred g p of 6X 103. (CH X employmg a 3 :1 molar ratio of oxrme to halide. For the unsaturated member (Formula III) the survival 5 2322 3?ig i il i z ri v illi ill 1 1 2 gg g g gi fig lfififzg g ggg ii 3 25 5 1 3 fif gig %mega-haloipropyl quaterlnarg salt in a 1.5:1 molar ratio. gave complete survival (ratios of 4/4 and 6/6) for anigf s j izii g g f the pyridine oxime and mals challenged 'by VX, how v 7 halide was dissolved in sufficient ethanol and refluxed for erticetgmpounds of Formula IVa showed anomalous propthe period of time specified in Table Procedure B.A mixture of the oxirne and halide was They g rafactlvatlon ,rates whlch were (but dissolved in about 100 ml. of ethanol and heated in a 200 ratlos Whlch j h 1gh as cqmpared to as 20 ml. capped pressure bottle (carbonated beverage type) for shown by Table X bemg bromlde' the length of time specified. The reaction mixtures were T ble 6 cooled to room temperature and the product removed by filtration. In several instances it was necessary to add Eeactiva surviyal absolute ether to effect complete precipitation. The prod- R g g ucts were recrystallized from ether. This procedure was an usually employed because of its simplicity.

TABLE 9 gives the procedure, yields and melting points 28532:::::::::::::::::::::::::::::::::::::::: 235E118 iii for representative compounds- 1 Table .9

The compounds of group IVb, which are closely related Substituents Melting to those of IVa, were somewhat less effective. When X", o a p Yield or ewas bromide the compound had the following proper- Halide R g2; ga s? ties: Reactivation rate constant 2x10 survival ratio 5 Point, (GB)2/4.

The compounds of group V were another group in I Br 1 300m which the results of therapeutic treatment against GB I Br 2; t2 3% were better as compared to 2-PAM then the reactivation B E 3 32-8 rate constants would suggest, as shown by the following 4.0 f: oi 13:68 70 300m table being bmmideiZ::::::::: 35 128 33 333% Z IVb Br B, 20 83.5 248-251m V Br B, 69 43 230-231 at V Br B, 64 10 223-226 (I R"EN+- i r l g e l t a t i b 45 Constant (GB) VTTT Br B, 90 16 201-203 11 N+- 1.3 10 4/4 Further details regarding the preparation and properties of certain of our compounds are given in the following publications: by us and our associates:

Pyridine Aldoximes by Edward J. Poziomek, Brennie E. Hackley, I12, and George M. Steinberg, Journal of a 4/4 Organic Chemistry, vol. 23, pp. 714-717, (May 1958); and Chemotherapeutic Efiectiveness of Trimethylene Bis (4-Formyl Pyridinium Bromide) Dioxime in Anticholin- The compounds of Formulas VI, VII and VIII, While l l by Edmunfi PP and being of dilferent structure are alike in exhibiting reactiva- Yates of the Soclety for Expenmental tion rate constants which are very low as compared to Y Pages i (May 1958)- 2 PAM but giving high survival ratios as shown (by Table These articles are to be considered lncorporated by ref- 8, X'" being bromide in each case. 111 this speclficauon- Whlle we have shown a number of specific examples of Table 8 compounds and their use, it will be obvious that various changes can be made without departing from our inven- Formllla gg ggg i g g tion, which is defined by the following claim.

We claim: W m 4/4 A 1,1'-(2-butenylene) bis (4 formylpyr1dinium) halide VII: Negligible. 4 4 dIOXIme- VIIL- 67 4/4 V No References Cited. 

